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Antirestriction Protein ArdB (R64) Interacts with DNA

Biochemistry. Biokhimiia. 2020.
Манухов И. В., Кудрявцева А. А.

Abstract—The antirestriction ArdB protein inhibits the endonuclease activity of type I restriction/modification (RM) systems in vivo; however, the mechanism of inhibition remains unknown. In this study, we showed that recombinant ArdB from Escherichia coli cells copurified with DNA. When overexpressed in E. coli cells, a portion of ArdB protein formed insoluble DNAfree aggregates. Only native ArdB, but not the ArdBΔD141 mutant lacking the antirestriction activity, copurified with DNA upon anionexchange and affinity chromatography or total DNA isolation from formaldehydetreated cells. These observations confirm the hypothesis that ArdB blocks DNA translocation via the R subunits of the R2M2S complex of type I RM enzymes

Language: English
DOI
Keywords: antirestriction
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ArdB, ArdA, and Ocr proteins inhibit the endonuclease activity of the type I restriction-modification enzymes (RMI). In this study, we evaluated the ability of ArdB, ArdA, and Ocr to inhibit different subtypes of Escherichia coli RMI systems (IA, IB, and IC) as well as two Bacillus licheniformis RMI systems. Furthermore we explored, the antirestriction activity of ArdA, ArdB, and ...
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Added: October 19, 2022
The Importance of C-Terminal Aspartic Acid Residue (D141) to the Antirestriction Activity of the ArdB (R64) Protein
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Added: October 19, 2022
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